Studies of the Biogenic Amine Transporters. 12. Identification of Novel Partial Inhibitors of Amphetamine-Induced Dopamine Release

摘要

Previous studies identified partial inhibitors and allosteric modulators of 5-HT ([5-amino-3-(3,4-dichlorophenyl)-1,2-dihydropyrido[3,4-b]pyrazin-7-yl]carbamic acid ethyl ester [SoRI-6238], 4-(2-[bis(4-fluorophenyl)methoxy]ethyl)-1-(2-trifluoromethyl-benzyl)-piperidine [TB-1-099]) and dopamine transporters N-(Diphenylmethyl)-2-phenyl-4-quinazolinamine, [SoRI-9804]). We report here the identification of three novel allosteric modulators of the dopamine transporter (N-(2,2-Diphenylethyl)-2-phenyl-4-quinazolinamine [SoRI-20040], N-(3,3-Diphenylpropyl)-2-phenyl-4-quinazolinamine [SoRI-20041], [4-Amino-6-[(diphenylmethyl)amino]-5-nitro-2-pyridinyl]carbamic acid ethyl ester, [SoRI-2827]). Membranes were prepared from HEK cells expressing the cloned human dopamine (hDAT) transporter. [¹²⁵I]RTI-55 binding and other assays followed published procedures. SoRI-20040, SoRI-20041 and SoRI-2827 partially inhibited [¹²⁵I]RTI-55 binding with EC50 values ranging from ∼1.4 μM to 3 μM and EMAX values decreasing as the [¹²⁵I]RTI-55 concentrations increased. All three compounds decreased the [¹²⁵I]RTI-55 Bmax and increased the apparent Kd in a manner well described by a sigmoid dose-response curve. In dissociation rate experiments, SoRI-20040 (10 μM) and SoRI-20041 (10 μM), but not SoRI-2827 (10 μM), slowed the dissociation of [¹²⁵I]RTI-55 from hDAT by ∼30%. Using rat brain synaptosomes, all three agents partially inhibited [³H]dopamine uptake with EC50 values ranging from 1.8 μM to 3.1 μM and decreased the VMAX value in a dose-dependent manner. SoRI-9804 and SoRI-20040 partially inhibited amphetamine-induced DAT-mediated release of [³H]MPP+ from rat caudate synaptosomes in a dose-dependent manner. Viewed collectively, we report several compounds that allosterically modulate hDAT binding and function, and identify novel partial inhibitors of amphetamine-induced dopamine release.